Question

In the dna isolation process, ________ is used to break down the protein complexes and allow the dna molecules to easily precipitate.

Answer 1

In the DNA isolation process, ice cold 95% ethanol is used to break down the protein complexes and allow the DNA molecules to easily precipitate.

What is separation and isolation of DNA fragments?

• The cutting of DNA by restriction endonucleases results in the fragments of DNA. These fragments can be separated by a technique known as gel electrophoresis.
• Since, DNA fragments are negatively charged molecules they can be separated by forcing them to move towards the anode under an electric field through a medium/matrix.
• The most commonly used matrix is agarose which is a natural polymer extracted from sea weeds.
• The DNA fragments separate (resolve) according to their size through sieving effect provided by the agarose gel. Hence, the smaller the fragment size, the farther it moves.
• The separated DNA fragments can be visualised only after staining the DNA with a compound known as ethidium bromide followed by exposure to UV radiation (you cannot see pure DNA fragments in the visible light and without staining). We can see bright orange coloured bands of DNA in a ethidium bromide stained gel exposed to UV light.
• The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece. This step is known as elution.
• The DNA fragments purified in this way are used in constructing recombinant DNA by joining them with cloning vectors.

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